Multiscale Analysis of Extracellular Matrix Remodeling in the Failing Heart

Abstract

Rationale: Cardiac extracellular matrix (ECM) comprises a dynamic molecular network providing structural support to heart tissue function. Understanding the impact of ECM remodeling on cardiac cells during heart failure (HF) is essential to prevent adverse ventricular remodeling and restore organ functionality in affected patients. Objective: We aimed to (i) identify consistent modifications to cardiac ECM structure and mechanics that contribute to HF and (ii) determine the underlying molecular mechanisms. Methods and Results: We first performed decellularization of human and murine ECM (dECM) and then analyzed the pathological changes occurring in dECM during HF by atomic force (AFM), two-photon microscopy, high-resolution 3D image analysis and computational fluid dynamics (CFD) simulation. We then performed molecular and functional assays in patient-derived cardiac fibroblasts (CFs) based on YAP-TEAD mechanosensing activity and collagen contraction assays. The analysis of HF dECM resulting from ischemic (IHD) or dilated cardiomyopathy (DCM), as well as from mouse infarcted tissue, identified a common pattern of modifications in their 3D topography. As compared to healthy heart, HF ECM exhibited aligned, flat and compact fiber bundles, with reduced elasticity and organizational complexity. At the molecular level, RNA sequencing of HF CFs highlighted the overrepresentation of dysregulated genes involved in ECM organization, or being connected to TGFß1, Interleukin-1, TNF-alpha and BDNF signaling pathways. Functional tests performed on HF CFs pointed at mechanosensor YAP as a key player in ECM remodeling in the diseased heart via transcriptional activation of focal adhesion assembly. Finally, in vitro experiments clarified pathological cardiac ECM prevents cell homing, thus providing further hints to identify a possible window of action for cell therapy in cardiac diseases. Conclusions: Our multi-parametric approach has highlighted repercussions of ECM remodeling on cell homing, CF activation and focal adhesion protein expression via hyper-activated YAP signaling during HF.

The Role of Tenascin C in Cardiac Reverse Remodeling Following Banding–Debanding of the Ascending Aorta

“…A recent publication by Perestrelo et al [25] added additional information to the puzzle of ECM remodeling. In their clinical and experimental decellularized samples of failing myocardium, they identified a common pattern of aligned, flat, and compact fiber bundles with reduced elasticity and organizational complexity.…”

Bearing My Heart: The Role of Extracellular Matrix on Cardiac Development, Homeostasis, and Injury Response

“…A versatile decellularization protocol working efficiently on distinct organs and also on the same tissue but on different ontogenetic stages (young and adult tissues) or health status has been recently reported (Silva et al, 2016(Silva et al, , 2019Garlikova et al, 2017;Pinto et al, 2017). This approach mitigates differences resultant from the application of distinct decellularization protocols, allowing fair comparisons on ECM composition and functional alterations across tissues, age, and normalcy vs. disease status (Perestrelo et al, 2020). Decellularization can be performed on tissues/organs or on cells cultured in vitro as monolayers or aggregates (Beacham et al, 2007;Nair et al, 2008).…”

The mechanical regulation of RNA binding protein hnRNPC in the failing heart

“…Thus far we have shown that cardiac ECM pathological remodeling induces hnRNPC shuttling to the active sites of translation at the sarcomere. Pathological remodeling is a complex phenomenon modifying the chemical composition and the mechanical properties of cardiac ECM 5,23 . We tried to disentangle the role of mechanical cues in hnRNPC localization.…”